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FITC anti-mouse CD182 (CXCR2) Antibody
149607
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I am interested in expression of CXCR2, which is expressed on neutrophils and is important for neutrophil migration. I am specifically interested in neutrophil recruitment to the lung. For the shown experiment I titrated the CXCR2 antibody on naive lungs. Without inflammation (in naive mice), neutrophils are only present in the capillaries of the lung. I confirmed that neutrophils were in capillaries with intravenous staining of CD45. Naive lungs were dissociated, digested with collagenase, and red blood cells were lysed. Leukocytes were stained with neutrophil and monocyte markers along with anti-CXCR2-FITC. Anti-CXCR2 was titrated in samples. Data show gated capillary neutrophils and inflammatory (Ly6C high) monocytes (which don't express CXCR2, as a negative control).
Flow Cytometry
Murine lung leukocytes (neutrophils in capillaries)
Dilutions (as shown) along with other marker antibodies; 30 minutes at 4 C.
Buffer contained BSA and FcBlock
None
Fluoresecence in flow cytometry
From my experience, staining cytokine receptors require higher concentrations of antibody. CXCR2 is expressed on circulating neutrophils and shown here lung capillary neutrophils do stain for CXCR2. Monocytes, which do not highly express CXCR2, don't stain with this antibody indicating that there isn't much off target staining. Of the titrations, 1:20 worked the best and showed definite positive staining on neutrophil, which are know to express CXCR2 on their surface.
N/A
Stains well
Requires higher amounts of antibody for staining
Would recommend this product for investigating CXCR2 expression.